Salmonella Typhimurium Research Articles

Applied Research Note: Combination of probiotic and prebiotic impacts Salmonella Enteritidis infection in layer pullets

Mitigation of food pathogens such as Salmonella continues to be a focus in poultry production, whether on shell eggs or meat products. No single approach is fully able to eliminate the risk of Salmonella contamination, but rather comprehensive programs are used in a multi-faceted approach. Targeted in-feed additives are a component of a food safety program and may help to reduce Salmonella infection. This study was aimed to evaluate the efficacy of a probiotic and prebiotic combination feed-additive product to reduce Salmonella Enteritidis (S.E.) infection in layer pullets. A total of 192 day-old pullets were placed in cages in two treatments (Control and pre-probiotic combination product at 1.5 lb/ton/of complete feed). Birds were challenged with 1.8 × 10 9 colony forming units (CFU) per bird of S.E. on Day 21 of the trial. Twenty-four birds from each treatment were sampled 6, 10, 14 and 18 days post infection (dpi). Cecal pouches were collected for enumeration of S.E. using plate counting technique. S.E. counts were numerically lower in Treatment 2 compared to Treatment 1 at 14 and 18 dpi, as Salmonella shedding in the course of infection was declining. Mean S.E. count in the ceca of pullets in Treatment 2 was 0.54 log 10 lower than that of Treatment 1 (3.36 log10) at 18 dpi. Samples below the detection limit of the counting method were significantly (p=0.021) higher in 10/24 (41.7%) of the pullets in Treatment 2 compared to 3/24 (12.5%) Treatment 1 at 18 dpi. Therefore, addition of the dietary pre- and probiotic combination product resulted in lower S.E. shedding in layer pullets challenged with S.E.

Efficacy of Oral Azithromycin in Treatment of Uncomplicated Enteric Fever in Children

Background: Enteric fever also known as (typhoid or paratyphoid fever) is a common systemic infection with increased incidence of morbidity and mortality with salmonella typhi and paratyphi. Azithromycin has shown better outcomes in children with uncomplicated disease. Objective: To determine the efficacy of Azithromycin in treatment of uncomplicated enteric fever in children. Study Design: Descriptive case series study Place and Duration of Study: Department of Pediatrics, Sir Ganga Ram Hospital Lahore, Pakistan from 21st August 2015 to 20th February 2016. Methodology: One hundred and sixty five cases were included. Patients received capsule/suspension of Azithromycin (10 mg/kg/day) OD for 7 days. Treatment was labelled as effective if both clinical and microbiological cure were observed. Results: Patients ranged between 2-12 years of age. Mean age of the patients was 6.45±2.39 year. There were 85 males (51.5%) and 80 females (48.5%) in this study. Mean duration of fever was 6.97±1.37 days. Clinical cure, microbiological cure and efficacy was 93.9%. Conclusion: Azithromycin to be effective and safe in treatment of enteric fever. We observed high cure rate of 93.9% without any serious side effects leading to the conclusion that azithromycin is an effective oral agent for treatment for enteric fever. Keywords: Enteric Fever, Azithromycin, Salmonella Typhi/Paratyphi

Detection of salmonella typhi transmitted by housefly in patient's blood in al-Diwaniyah province, Iraq

Salmonella typhi is a gram- negative bacterium, it is causes disease called typhoid fever. House flies are play important role in transmission of Salmonella. This study aimed to detect Salmonella typhi transmitted by house flies in patient's blood in Al-Diwaniyah province of Iraq. This study was collected 30 blood samples from patient's in Al-Diwaniyah teaching hospital from November 2021 until to end March 2022 from both sexes (17 males and 13 females) and age groups ranged between (10-50) years. The results of study revealed that high prevalence of disease in males 17(56.66%), but revealed low prevalence of disease in females13(43.33%). According to age groups the result was recorded high incidence of disease in age group (10-20) (40%), while it was recorded low incidence of disease in age group (40-50) (10%). In current study, 10 serum samples out of 30 samples were detected 6(60%) positive samples for IgM, whereas 4(40%) positive samples for IgG in rapid test of Salmonella typhi. 

Weighted gene co-expression network analysis identifies potential regulators in response to Salmonella Enteritidis challenge in the reproductive tract of laying ducks

Salmonella Enteritidis (SE) is a zoonotic and vertically transmitted pathogen, often colonized in the reproductive tract of adult poultry, which can result in direct contamination of eggs and threaten human health. Previous studies have revealed that some pattern recognition receptors and resistance genes were involved in regulating immune responses to SE invasion in birds. However, the role of these immune response genes was not independent, and the interactions among the genes remained to be further investigated. In this study, SE burden and colonization were determined in reproductive tissue after the ducks were SE-infected, and RNA-sequencing was performed to construct co-expression networks by weighted gene co-expression network analysis (WGCNA). The result showed that SE could be isolated from 22% of infected-birds in any segment of the reproductive tract and the SE was readily colonized in the stroma, small follicle, isthmus, and vagina of the reproductive tracts in morbid ducks. The top central, highly connected genes were subsequently identified three specific modules in the above four tissues at the defined cut-offs (P<0.01), including 60 new candidate regulators and 125 transcription factors. Moreover, those 185 differentially expressed genes (DEGs) in these modules were co-expressed. Moreover, the hub genes (TRAF3, CXCR4 and IL13RA1) were identified to act with many other genes through immune response pathways including NF-kappaB, Toll-like receptor, steroid biosynthesis, and p53 signaling pathways. These data provide references that will understand the immune regulatory relationships during SE infection, but also assist in the breeding of SE-resistant lines through potential biomarkers.

Transcriptome analysis of the spleen of heterophils to lymphocytes ratio-selected chickens revealed their mechanism of differential resistance to Salmonella

Salmonella is one of the most common food-borne pathogens and its resistance in chicken can be improved through genetic selection. The heterophils/lymphocytes (H/L) ratio in the blood reflects the immune system status of chicken. We compared the genome data and spleen transcriptomes between the H/L ratio-selected and non-selected chickens, after Salmonella infection, aiming to identify the key genes participating in the antibacterial activity in the spleen. The results revealed that, the selected population had stronger (P<0.05) liver resistance to Salmonella typhimurium (ST) than the non-selected population. In the selected and non-selected lines, the identified differentiation genes encode proteins involved in biological processes or metabolic pathways that included the TGF-beta signaling pathway, FoxO signaling pathway, and Salmonella infection pathway. The results of the analysis of all identified differentially expressed genes (DEGs) of spleen revealed that the G protein-coupled receptor (GPCR) and insulin-like growth factor (IGF-I) signaling pathways were involved in the Salmonella infection pathway. Integrated analysis of DEGs and FST (fixation index), identified candidate genes involved in Salmonella infection pathway, such as GPR39, NTRK2, and ANXA1. The extensive genomic changes highlight the polygenic genetic of the immune response in these chicken populations. Numerous genes related to the immune performance are differentially expressed in the selected and non-selected lines and the selected lines has a higher resistance to Salmonella.

Fluorescent aptasensor for detection of live foodborne pathogens based on multicolor perovskite-quantum-dot-encoded DNA probes and dual-stirring-bar-assisted signal amplification

In this study, a fluorescent (FL) aptasensor was developed for on-site detection of live Salmonella typhimurium (S.T.) and Vibrio parahaemolyticus (V.P.). Complementary DNA (cDNA) of aptamer (Apt)-functionalized multicolor polyhedral oligomeric silsesquioxane-perovskite quantum dots (cDNA-POSS-PQDs) were used as encoded probes and combined with dual-stirring-bar-assisted signal amplification for pathogen quantification. In this system, bar 1 was labeled with the S.T. and V.P. Apts, and then bar 2 was functionalized with cDNA-POSS-PQDs. When S.T. and V.P. were introduced, pathogen-Apt complexes would form and be released into the supernatant from bar 1. Under agitation, the two complexes reached bar 2 and subsequently reacted with cDNA-POSS-PQDs, which were immobilized on MXene. Then, the encoded probes would be detached from bar 2 to generate FL signals in the supernatant. Notably, the pathogens can resume their free state and initiate next cycle. They swim between the two bars, and the FL signals can be gradually enhanced to maximum after several cycles. The FL signals from released encoded probes can be used to detect the analytes. In particular, live pathogens can be distinguished from dead ones by using an assay. The detection limits and linear range for S.T. and V.P. were 30 and 10 CFU/mL and 102–106 CFU/mL, respectively. Therefore, this assay has broad application potential for simultaneous on-site detection of various live pathogenic bacteria in water.

A multi-jurisdictional outbreak of Salmonella Typhimurium infections linked to backyard poultry-Australia, 2020.

Zoonotic salmonellosis can occur either through direct contact with an infected animal or through indirect contact, such as exposure to an infected animal's contaminated environment. Between May and August 2020, a multi-jurisdictional outbreak of Salmonella Typhimurium (STm) infection due to zoonotic transmission was investigated in Australia. In total, 38 outbreak cases of STm with a median age of 5 years were reported. Epidemiological investigation showed contact with live poultry to be a common risk factor with most cases recently purchasing one-week old chicks from produce/pet stores. Traceback investigation of cases identified 25 product/pet stores of which 18 were linked to a single poultry breeder farm. On farm environmental sampling identified the same STm genotype as identified in cases. Whole genome sequencing of both environmental and human outbreak isolates found them to be highly related by phylogenetic analysis. This investigation describes the first documented widespread zoonotic salmonellosis outbreak in Australia attributed to backyard poultry exposure and identified potential risk factors and prevention and control measures for future outbreaks. Prevention of future outbreaks will require an integrated One Health approach involving the poultry industry, produce/pet store owners, animal healthcare providers, public health and veterinary health agencies and the public.

Effects of microencapsulated essential oils on growth performance and biomarkers of inflammation in broiler chickens challenged with salmonella enteritidis

Feed ingredients are the potential sources, which can be contaminated with Salmonella. Therefore, the alternative strategies should be found for controlling pathogenic microbes and promoting the performance of broiler chicks. Although antibiotics have been used for the treatment of the infection induced by microbes, the use of which faces with some limitations such as bacteria resistance. The antibiotic substitute should be natural with no residue in poultry meat. Compounds such as phytobiotics can be incorporated into the diet for enhancing the production of poultry with no concern about the appearance of antibiotic-resistant microbial species and remaining the residue in the meat. This study aims to synthesize the microcapsules preserving the mixture of essential oils including thyme, peppermint, savory, and black pepper by using spray drying technique and to evaluate the growth performance, antioxidant status, immune response, and gut health parameters in broiler chickens challenged by salmonella enteritidis. The mixture of essential oils including thyme (50%), summer savory (25%), peppermint (12.5%) and black pepper seed (12.5%) was encapsulated in the protecting wall materials (20% WPC, 25% MS and 55% MD) for forming the essential oil-loaded spherical microcapsules with particle size of 323 nm and encapsulation efficiency of 96.2%. Thymol, carvacrol, p-cymene, γ-terpinene, and menthol existed in the core of the microcapsules as the main bioactive compounds. The microcapsules possessed the antioxidant and antibacterial activities, and its dietary application as the feed additive at the concentrations of 0.5, 1 and 2 kg/ton in broiler chickens challenged by S. enteritidis improved the final weight, total feed intake, FCR, body antioxidant status, ileal morphostructure, intestinal microbial population and regulated the antioxidant and inflammation genes in ileal tissue and upregulated the expression of occludin and SOD genes.

ISOLATION AND IDENTIFICATION OF SALMONELLA SPP FROM CHICKEN MEAT IN KURDISTAN REGION

Salmonella is a bacterial diseases that caused by many strains of salmonella. Salmonellosis is a disease occur in human and chicken, salmonellosis in poultry sometimes causes economic losses, also international trade can be affected if the salmonella presents in poultry products or poultry meat as in this case the infection can transmit to humans. Salmonella is a considered the most important and serious pathogen of bacteria that responsible of food borne infection all over the world. In Kurdistan region, Erbil city the common shapes of poultry are broilers and backyard. This study was done to inform the impact of salmonella epidemic chicken and the prevalence of salmonella's genetic types and serotypes among broilers and raw meat of chickens in Kurdistan, Iraq. Samples were collected from about fifty broiler chickens that suffered from diarrhea, fifty raw chickens meat and "30" patients that had diarrhea and signs of food poisoning. The serological identification of salmonella after isolation was "58.33%" salmonella enteritidis and "41.66%" salmonella typhimurium. Serotype salmonella enteritidis antigenic formula has "O" somatic antigen "1, 9, 12" and flagellar antigen "H", while the serotype salmonella typhimurium had the "O" somatic antigen "1, 4". The main infection source of humans includes; meat products that comes from consumption of contaminated meat of chicken. Scientifically, the control programs of impact of salmonella control are based on general procedures of hygiene on the campylobacter prevalence at the holding in broiler flocks and at the end of the process of slaughter on the broiler meat.

Effect of enzyme-modified yeast products on Salmonella Enteritidis colonization in different organs of laying hens

A study was conducted to assess the effectiveness of yeast-derived products in the recovery of Salmonella Enteritidis (SE) in White Leghorns. A total of thirty-two hens at 38 weeks of age were challenged both orally (OR) and intracloacally (IC) with nalidixic acid resistant SE (SE NAR ). Hens were individually housed in conventional wire laying cages and randomly allocated to four treatments (T) with eight hens per replicate: 1) No SE NAR challenge (T1; basal diet), 2) SE NAR challenge (T2; basal diet), 3) SE NAR challenge: basal diet with a blend of Saccharomyces- Torula yeasts and yeast cell walls (T3; intact yeast), and 4) SE NAR challenge: basal diet with enzymatically-modified yeast products blend (T4; enzyme-modified yeast). The excreta samples were 100% positive for SE NAR at 3 days post-infection (dpi) in T2, T3, and T4. By 6 dpi, excreta samples were 87, 62 and 37% positive for T2, T3, and T4, respectively. Hens were euthanized at 7 dpi to harvest liver with gall bladder (L/GB), spleen, ovaries, and ceca. The recovery of SE NAR in L/GB was 75, 25 and 37% in T2, T3, and T4 (P < 0.05), whereas that of the spleen was 62, 25 and 37% in T2, T3 and T4 (P < 0.05), respectively. Treatments 3 and 4 reduced SE NAR counts (cfu/g) in the ceca from 4.3 log 10 to 2.7 and 1.95 log 10 , respectively, compared to T2 ( P < 0.05). The cecal count of SE NAR was 1.6 log 10 lower in T3 and 2.3 log 10 lower in T4 compared to T2. Both yeast derived products reduced Salmonella in internal organs and the ceca of laying hens. Enzymatically-modified yeast (T4) was better than its intact counterpart (T3) in reducing Salmonella counts in the ceca.

Biosynthesis of Salmonella enteritidis O antigen-based glycoproteins

Salmonella enteritidis (SE) has been recognized as an important zoonotic pathogen, and the prevention and control of salmonellosis has long been a conundrum. However, glycoconjugate vaccines seem to be a promising solution. Glycoproteins are conventionally synthesized by chemical cross-linking which features complex procedure and cost-intensiveness. Therefore, a stable biosynthesis method at lower cost is in urgent need. For the biosynthesis of SE O-antigen-based glycoproteins, we used CRISPR/Cas9 to develop the waaL-deleted SE strain ∆waaL. The synthesis of lipopolysaccharide (LPS) was detected based on silver staining. Circular polymerase extension cloning (CPEC) was employed to construct the plasmids expressing glycosyltransferase PglL, recombinant Pseudomonas aeruginosa exotoxin A (rEPA), and cholera toxin B subunit (CTB). Meanwhile, PilES45-K73 glycosylation motif was added to the N-terminal and C-terminal of rEPA and CTB, respectively. The recombinant plasmids were transformed into SE ∆waaL. After induction, the synthesis of glycoprotein was verified by Western blotting and the synthesized glycoprotein was purified by Ni-NTA column. The results showed that waaL deletion blocked the LPS synthesis of SE, and that rEPA and CTB proteins were expressed in SE ∆waaL. In addition, obvious glycosylation occurred to rEPA and CTB when PglL was expressed, and the glycosylated part was SE O antigen polysaccharide. In summary, after waaL deletion in SE, PglL can transfer its own O antigen polysaccharides (OPS) to the carrier proteins rEPA and CTB, resulting in OPS-rEPA and OPS-CTB glycoproteins. The result lays a basis for the biosynthesis of SE glycoprotein.

Study of some microbial infections of recurrent miscarriage in women

Background: some microbial infections can be passed to the fetus before or during birth and damage the fetus or cause a miscarriage, like infection with ToRCH agents and salmonella enterica. The current study aims to investigate the extent of the involvement of salmonella enteria serovars, proteus spp. and Brucella spp. in recurrent miscarriage in women during first trimester. Material and method: The 50 blood samples were collected from women age rage (19-39), who had repeated miscarriages during the first trimester during periods extended from October 2021 to February 2022,. The study control includes the 20 fertile women with no miscarried history. Two tests were conducted which are ToRCH IgM and febril antigen slid test for identification infectious agents associated with pregnancy loss. Results: The present results showed that the most frequent recurrent miscarriage (RM) was in the age group 35-39 years at a percentage 34%, while the lowest abortion rate was 18% at the age group of 25-29 years.

Genome-wide alternative splicing profile in the posterior kidney of brown trout (Salmo trutta) during proliferative kidney disease

BackgroundThe cnidarian myxozoan parasite Tetracapsuloides bryosalmonae causes chronic proliferative kidney disease (PKD) in salmonids. This parasite is a serious threat to wild and cultured salmonids. T. bryosalmonae undergoes intra-luminal sporogonic development in the kidney of brown trout (Salmo trutta) and the viable spores are released via urine. We investigated the alternative splicing pattern in the posterior kidney of brown trout during PKD.ResultsRNA-seq data were generated from the posterior kidney of brown trout collected at 12 weeks post-exposure to T. bryosalmonae. Subsequently, this data was mapped to the brown trout genome. About 153 significant differently expressed alternatively spliced (DEAS) genes, (delta PSI = 5%, FDR P-value < 0.05) were identified from 19,722 alternatively spliced events. Among the DEAS genes, the least and most abundant alternative splicing types were alternative 5′ splice site (5.23%) and exon skipping (70.59%), respectively. The DEAS genes were significantly enriched for sodium-potassium transporter activity and ion homeostasis (ahcyl1, atp1a3a, atp1a1a.1, and atp1a1a.5). The protein-protein interaction network analysis enriched two local network clusters namely cation transporting ATPase C-terminus and Sodium/potassium ATPase beta chain cluster, and mixed inclusion of Ion homeostasis and EF-hand domain cluster. Furthermore, the human disease-related salmonella infection pathway was significantly enriched in the protein-protein interaction network.ConclusionThis study provides the first baseline information about alternative splicing in brown trout during PKD. The generated data lay a foundation for further functional molecular studies in PKD - brown trout infection model. The information generated from the present study can help to develop therapeutic strategies for PKD in the future.

Physiological and Proteomic Analysis Responsive Mechanisms for Salt Stress in Oat.

Oat is considered as a moderately salt-tolerant crop that can be used to improve saline and alkaline soils. Previous studies have focused on short-term salt stress exposure, and the molecular mechanisms of salt tolerance in oat have not yet been elucidated. In this study, the salt-tolerant oat cultivar Vao-9 and the salt-sensitive oat cultivar Bai5 were treated with 6 days of 0 and 150 mmol L−1 salt stress (nNaCl:nNa2SO4 = 1:1). Label-Free technology was then used to analyze the differentially expressed proteins in leaves under 0 and 150 mmol L−1 salt stress. The obtained results indicated that total of 2,631 proteins were identified by mass spectrometry in the four samples. The salt-tolerant cultivar Vao-9 mainly enhances its carbohydrate and energy metabolism through the pentose and glucuronate interconversions, and carbon fixation pathways in prokaryotes, thereby reducing the damage caused by salt stress. In addition, the down-regulation of ribosomes expression and the up-regulated expression of HSPs and CRT are all through the regulation of protein synthesis in response to salt stress. However, GABA metabolism presents a different synthesis pattern in Bai5 and Vao-9. The main KEGG function of differential expressed protein (DEP) in Bai5 is classified into protein processing in the endoplasmic reticulum, estrogen signaling pathway, antigen processing and presentation, longevity regulating pathway-multiple species, arginine and proline metabolism, beta-alanine metabolism, vitamin B6 metabolism, salmonella infection, chloroalkane and chloroalkene degradation, and limonene and pinene degradation. Moreover, the main KEGG functions of DEP in Vao-9 are classified as ribosome and carbon fixation pathways in prokaryotes, pentose and glucuronate interconversions, GABA ergic synapse, and taurine and hypotaurine metabolism. The results obtained in this study provide an important basis for further research on the underlying mechanisms of salt response and tolerance in oat and other plant species.

Research Note: In-feed Bacteriophage does not impact efficacy of live Salmonella vaccine

Dietary bacteriophages potentially can serve as a step to reduce Salmonella contamination of feed through direct lysis of the bacteria. However, poultry producers commonly vaccinate with live Salmonella vaccines, which could potentially be lysed by dietary bacteriophages. The objective of this study was to evaluate if dietary bacteriophages impacted the colonization of a live Salmonella vaccine. A total of 210 day-of-hatch Ross male broiler chicks were divided into 3 treatments consisting of 2 replicate per treatment. Each replicate contained 35 birds. T1 was the challenge control, given no Salmonella vaccine, T2 was challenged and given Salmonella vaccine and T3 was challenged, given Salmonella vaccine as well as dietary bacteriophage. Salmonella vaccine was administered day of hatch. On d 3, four birds/pen were sampled for Salmonella vaccine colonization of ceca and liver/spleen. The remaining birds were challenged with 5 × 107 CFU of nalidixic acid- resistant Salmonella enteritidis (S.E.). On d 28, ten birds/replicate were sampled via cloaca swabs to culture for S.E. On d 42, the trial was terminated, birds were weighed, and performance was calculated. In addition, 15 birds/replicate were sampled for cecal cultures of S.E.On d 3, T1 had 0% vaccine strain isolated, and significantly lower (P = 0.009) cecal prevalence compared with T2 (75%) and T3 (38%) being intermediate. T1 (0%) had significantly lower liver/spleen vaccine strain prevalence (P = 0.002) compared with T3 (88%) and T2 (63%) being intermediate. No significant differences (P > 0.05) were observed among treatments in Salmonella prevalence in d 28 cloacal swabs. No significant differences (P > 0.05) were observed in d 42 cecal Salmonella prevalence between all treatments. No significant differences in bird weight were observed between treatments d 0 to 42 (P > 0.05). However, T2 and T3 had lower mortality and adjusted feed conversion ratio (FCR; P < 0.05) compared with T1. Therefore, the dietary bacteriophage did not interfere with colonization or protection afforded by the live Salmonella vaccine.

Vitamin B12 coordinates ileal epithelial cell and microbiota functions to resist Salmonella infection in mice.

Deprivation of vitamin B12 (VB12) is linked to various diseases, but the underlying mechanisms in disease progression are poorly understood. Using multiomic approaches, we elucidated the responses of ileal epithelial cells (iECs) and gut microbiome to VB12 dietary restriction. Here, VB12 deficiency impaired the transcriptional and metabolic programming of iECs and reduced epithelial mitochondrial respiration and carnitine shuttling during intestinal Salmonella Typhimurium (STm) infection. Fecal microbial and untargeted metabolomic profiling identified marked changes related to VB12 deficiency, including reductions of metabolites potentially activating mitochondrial β-oxidation in iECs and short-chain fatty acids (SCFAs). Depletion of SCFA-producing microbes by streptomycin treatment decreased the VB12-dependent STm protection. Moreover, compromised mitochondrial function of iECs correlated with declined cell capability to utilize oxygen, leading to uncontrolled oxygen-dependent STm expansion in VB12-deficient mice. Our findings uncovered previously unrecognized mechanisms through which VB12 coordinates ileal epithelial mitochondrial homeostasis and gut microbiota to regulate epithelial oxygenation, resulting in the control of aerobic STm infection.

Research Note: Effect of corn silage and alfalfa meal as alternative induced molt methods to improving Salmonella Enteritidis resistance in laying hens

This experiment was conducted to evaluate diets containing a high level of corn silage and alfalfa meal in inducing molt and reducing susceptibility to Salmonella Enteritidis (SE) colonization in laying hens. Thirty-two healthy hens were examined by cloacal swab samples to be free of Salmonella. Then they were weighed individually and distributed to 4 experimental groups containing 8 hens each, including Full-fed (control, FF); total feed withdrawal (positive control for molt induction, FW); 80% corn silage (CS) + 20% layer diet (CS80), and 80% alfalfa meal (AM) + 20% layer diet (AM80). The molting program was initiated at 71 wk of age. On d 4 of the experiment, all hens were inoculated with SE by oral gavage. All hens were first weighed at the ending molting period on d 10 and then euthanized by CO2 gas. The internal organs including the ovary, oviduct, liver, and spleen, were excised aseptically and weighed. Cloacal swab and feed samples at the beginning and organ samples (liver, ovary, spleen, and cecum) were collected from each hen at the end of the experiment and examined for SE colonies. Molted birds lost roughly 14 to 27%t of their body weight and had significantly lower organ weight and egg production compared to FF group (P < 0.05). No significant difference was observed in the number of days to zero egg production between molted treatments. The SE positive organs did not significantly differ between CS80 and AM80 with FF treatment. Treatment CS80 had the lowest crop pH and differed substantially from treatment FW. In conclusion, results indicate that using corn silage and alfalfa meal, can improve resistance to salmonella Enteritidis during molt inducing compared to traditional feed withdrawal.

Development of Multiplex-PCR Method to Detect Three Bacterial Species in Food and their Use in Food Inspection

Food contains several microorganisms that may cause illnesses and food poisoning in humans. Small numbers of microorganism contamination could result in rapid spoilage of food. The Centres for Disease Control and Prevention (CDC), USA estimates that 76 million people are affected by foodborne illnesses each year in the USA. Salmonella infections alone account for one billion dollars yearly in direct and indirect medical costs and more than 5,000 deaths. In Sudan, diarrhoeal disease was reported as the second major disease during the years from 2003 to 2007 (Annual health statistical report of the Federal Ministry of Health, Sudan). We aimed to develop a rapid molecular procedure for the detection of Escherichia coli, Shigella dysentery, and salmonella Typhiin food so as to minimize the public health hazard of food contamination. We used the Multiplex PCR method as rapid methods were tested for identification of Enterobacteriaceae species Escherichia coli as an indicator organism for food contamination and two strains of Enterobacteriaceae that causes food borne illness (namely Shigella dysentery and salmonella Typhi). The Multiplex PCR was performed to detect E. coli using Mdh primer pair, Salmonella Typhi using IpaB primer pair, and Shigella dysentery using IpaH1 primer pair. The sensitivity to detect E. coli, Salmonella Typhi, and Shigella dysentery in contaminated food in the concentration of the infective and the over infective doses were 100%, 96.3%, and 88.9% respectively for the three bacteria strains. There was no significant difference in the detection of the bacteria after incubation for 8 hours, 24 hours, or even without incubation period. There were no differences in the result of the samples that were contaminated artificially in laboratory and those obtained from the market. The Multiplex PCR method for identification of E. coli, Salmonella Typhi and Shigella dysentery was developed as a model for detection and risk assessment of the three bacteria in one program, and it is suitable for routine analysis.

Identification of differentially expressed genes associated with coronary in-stent restenosis by integrated bioinformatics approaches.

In-stent restenosis (ISR) is a common complication following percutaneous coronary intervention (PCI). Designing ISR-specific therapeutic targets is important for optimizing coronary stenosis therapy. This study aims to find new potential key genes related to ISR and the usefulness of differentially expressed genes (DEGs) as diagnostic biomarkers for ISR. Microarray datasets of peripheral blood was downloaded from the Gene Expression Omnibus (GEO) public database (GSE46560, n=11). The DEGs between the ISR group and the control were obtained through the limma package analysis of the R language. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were used to obtain the related signal pathways. STRING database was used to constructed a protein-to-protein interaction network with DEGs. Networkanalyst database was used to predict the target miRNA of the target gene. The area under the curve (AUC) value obtained from receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic effectiveness of the hub gene in distinguishing ISR patients from normal individuals. A total of 154 DEGs were identified between ISR group and control group, including 57 up-regulated and 97 down-regulated DEGs. GO enrichment analysis showed that genes were enriched in homeostasis, cell morphology and cell connection. KEGG pathway analysis shows that genes are mainly involved in human papillomavirus infection, salmonella infection, human T-cell leukemia virus type 1 infection, axon guidance and MAPK signaling pathway. CLTA, CAT, STAT5A, CD300A, CA1, NCF2, HBQ1, AHSP, SLC4A1 and EPB42 were identified as 10 hub genes. A total of 151 target miRNAs of 7 DEGs were obtained, and 149 mRNA-miRNA pairs were identified. ROC curve indicates that CA1, STAT5A and HBQ1 have high diagnostic value in ISR. CA1, STAT5A and HBQ1 are identified as specific related genes for ISR patients. These genes may provide new targets for the early identification and treatment of ISR.

A Dose-Response Investigation of a Micronized Porous Ceramic Particle to Improve the Health and Performance of Post-weaned Pigs Infected With Salmonella enterica Serotype Typhimurium

The objective was to investigate the effects of supplementing increasing concentrations of PowerGuard (PG), a micronized ceramic particle, to weaned pigs on health and performance following a Salmonella enterica serotype Typhimurium infection. Forty barrows were transported to the USDA facility in Liberty, TX, USA. Pigs were randomly assigned to one of five treatments (n = 8): (1) uninfected control (CON), no Salmonella typhimurium (ST) and no PG treatment; (2) infected control (ST), infected with ST but no PG treatment; (3) PG0.05, infected with ST and supplemented with 0.05% PG; (4) PG0.25, infected with ST and supplemented with 0.25% PG; and (5) PG0.50, infected with ST and supplemented with 0.5% PG. All pigs were enrolled at 21.5 ± 1.33 days of age and did not differ in initial BW (1.98 ± 0.09 kg). Pigs were anesthetized to insert temperature recording devices into the abdominal cavity. Pigs were offered feed and water ad libitum. Pigs in ST, PG0.05, PG0.25, and PG0.50 were infected orally with 1.75 × 107 colony-forming units of Salmonella typhimurium on day 7. Pig body weights and peripheral blood samples were collected on days 0, 7, 10, 14, and 21. Pigs were harvested on day 21 and ileum and liver samples were collected for histopathological analyses. There was no treatment difference for final BW (P ≥ 0.201). There was a tendency (P = 0.087) for a treatment difference in the fecal score; ST and PG0.50 had more loose fecal scores than CON and PG0.25. There was a treatment × time interaction for intraperitoneal temperature (P &amp;lt; 0.0001); PG0.05, PG0.25, and PG0.50 had attenuated febrile responses during the acute post-infection period compared with ST. There was a treatment × time interaction for total leukocyte counts (P = 0.007); PG treatments reduced leukocytosis post-infection compared with ST. Supplementing PG0.25 improved many health and performance variables when pigs were infected with Salmonella Typhimurium. Furthermore, supplementing PG0.05 attenuated the febrile response and many hematological variables. However, supplementing PG0.5 did not improve many aspects of health or performance. Therefore, supplementing PowerGuard between 0.05 and 0.25% of the diet may play a role in protecting weaned pigs from disease caused by Salmonella.